Located in the area between Paseo de la Reforma and Mahatma Gandhi Street within Chapultepec Park in Mexico City,[3] the museum contains significant archaeological and anthropological artifacts from Mexico's pre-Columbian heritage, such as the Stone of the Sun (or the Aztec calendar stone) and the Aztec Xochipilli statue.

In December 1940, the museum was divided again, with its historical collections being moved to the Chapultepec Castle, where they formed the Museo Nacional de Historia, focusing on the Viceroyalty of the New Spain and its progress towards modern Mexico.

The remaining collection was renamed the National Museum of Anthropology, focusing on pre-Columbian Mexico and modern day Mexican ethnography.

The construction of the building lasted 19 months, and was inaugurated on September 17, 1964, President Adolfo López Mateos, who declared: The Mexican people lift this monument in honor of the admirable cultures that flourished during the Pre-Columbian period in regions that are now territory of the Republic.

The museum's collections include the Stone of the Sun, giant stone heads of the Olmec civilization that were found in the jungles of Tabasco and Veracruz, treasures recovered from the Maya civilization, at the Sacred Cenote at Chichen Itza, a replica of the sarcophagal lid from Pacal's tomb at Palenque and ethnological displays of contemporary rural Mexican life.

It also has a model of the location and layout of the former Aztec capital Tenochtitlan, the site of which is now occupied by the central area of modern-day Mexico City.